Abstract
Microtissues (MT) are currently considered as a promising alternative for the fabrication of natural, 3D biomimetic functional units for the construction of bio-artificial substitutes by tissue engineering (TE). The aim of this study was to evaluate the possibility of generating mesenchymal cell-based MT using human umbilical cord Wharton’s jelly stromal cells (WJSC-MT). MT were generated using agarose microchips and evaluated ex vivo during 28 days. Fibroblasts MT (FIB-MT) were used as control. Morphometry, cell viability and metabolism, MT-formation process and ECM synthesis were assessed by phase-contrast microscopy, functional biochemical assays, and histological analyses. Morphometry revealed a time-course compaction process in both MT, but WJSC-MT resulted to be larger than FIB-MT in all days analyzed. Cell viability and functionality evaluation demonstrated that both MT were composed by viable and metabolically active cells, especially the WJSC during 4–21 days ex vivo. Histology showed that WJSC acquired a peripheral pattern and synthesized an extracellular matrix-rich core over the time, what differed from the homogeneous pattern observed in FIB-MT. This study demonstrates the possibility of using WJSC to create MT containing viable and functional cells and abundant extracellular matrix. We hypothesize that WJSC-MT could be a promising alternative in TE protocols. However, future cell differentiation and in vivo studies are still needed to demonstrate the potential usefulness of WJSC-MT in regenerative medicine.
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References
Achilli TM, Meyer J, Morgan JR (2012) Advances in the formation, use and understanding of multi-cellular spheroids. Expert Opin Biol Therapy 12:1347–1360. https://doi.org/10.1517/14712598.2012.707181
Al Madhoun A, Ali H, AlKandari S, Atizado VL, Akhter N, Al-Mulla F, Atari M (2016) Defined three-dimensional culture conditions mediate efficient induction of definitive endoderm lineage from human umbilical cord Wharton’s jelly mesenchymal stem cells. Stem Cell Res Therapy 7:165. https://doi.org/10.1186/s13287-016-0426-9
Alaminos M, Del Carmen Sanchez-Quevedo M, Munoz-Avila JI, Serrano D, Medialdea S, Carreras I, Campos A (2006) Construction of a complete rabbit cornea substitute using a fibrin-agarose scaffold. Investig Ophthalmol Vis Sci 47:3311–3317. https://doi.org/10.1167/iovs.05-1647
Atala A (2012) Regenerative medicine strategies. J Pediatric Surg 47:17–28. https://doi.org/10.1016/j.jpedsurg.2011.10.013
Berneel E, Philips C, Declercq H, Cornelissen R (2016) Redifferentiation of High-throughput generated fibrochondrocyte micro-aggregates: impact of low oxygen tension. Cells Tissues Organs https://doi.org/10.1159/000447509
Campos F et al (2016) Ex vivo characterization of a novel tissue-like cross-linked fibrin-agarose hydrogel for tissue engineering applications. Biomed Mater 11:055004. https://doi.org/10.1088/1748-6041/11/5/055004
Campos F et al. (2017) Generation of genipin cross-linked fibrin-agarose hydrogels tissue-like models for tissue engineering applications. Biomed Mater. https://doi.org/10.1088/1748-605X/aa9ad2
Carriel VGI (2009) Aplicación del método de bolque celular para evaluar la población de fibroblastos de mucosa oral en ingeniería tisular. Actual Med 94/2009:007–011
Carriel V, Garzon I, Alaminos M, Campos A (2011a) Evaluation of myelin sheath and collagen reorganization pattern in a model of peripheral nerve regeneration using an integrated histochemical approach. Histochem Cell Biol 136:709–717. https://doi.org/10.1007/s00418-011-0874-3
Carriel VS, Aneiros-Fernandez J, Arias-Santiago S, Garzon IJ, Alaminos M, Campos A (2011b) A novel histochemical method for a simultaneous staining of melanin and collagen fibers. J Histochem Cytochem 59:270–277. https://doi.org/10.1369/0022155410398001
Carriel V et al. (2012) Epithelial and stromal developmental patterns in a novel substitute of the human skin generated with fibrin-agarose biomaterials. Cells Tissues Organs 196:1–12 https://doi.org/10.1159/000330682
Carriel V et al (2013) Combination of fibrin-agarose hydrogels and adipose-derived mesenchymal stem cells for peripheral nerve regeneration. J Neural Eng 10:026022. https://doi.org/10.1088/1741-2560/10/2/026022
Carriel V, Alaminos M, Garzon I, Campos A, Cornelissen M (2014) Tissue engineering of the peripheral nervous system. Expert Rev Neurother 14:301–318. https://doi.org/10.1586/14737175.2014.887444
Carriel V et al (2015) In vitro characterization of a nanostructured fibrin agarose bio-artificial nerve substitute. J Tissue Eng Regen Med. https://doi.org/10.1002/term.2039
Carriel V, Garzon I, Campos A, Cornelissen M, Alaminos M (2017) Differential expression of GAP-43 and neurofilament during peripheral nerve regeneration through bio-artificial conduits. J Tissue Eng Regen Med 11:553–563. https://doi.org/10.1002/term.1949
Davies JE, Walker JT, Keating A (2017) Concise review: Wharton’s jelly: the rich, but enigmatic, source of mesenchymal stromal cells. Stem Cells Transl Med. https://doi.org/10.1002/sctm.16-0492
Dean DM, Napolitano AP, Youssef J, Morgan JR (2007) Rods, tori, and honeycombs: the directed self-assembly of microtissues with prescribed microscale geometries. FASEB J 21:4005–4012. https://doi.org/10.1096/fj.07-8710com
Dominici M et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317. https://doi.org/10.1080/14653240600855905
Donders R et al (2018) Human Wharton’s jelly-derived stem cells display a distinct immunomodulatory and proregenerative transcriptional signature compared to bone marrow-derived stem cells. Stem Cells Dev 27:65–84. https://doi.org/10.1089/scd.2017.0029
Edmondson R, Adcock AF, Yang L (2016) Influence of matrices on 3D-cultured prostate cancer cells’ drug response and expression of drug-action associated proteins. PloS One 11:e0158116. https://doi.org/10.1371/journal.pone.0158116
Fennema E, Rivron N, Rouwkema J, van Blitterswijk C, de Boer J (2013) Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol 31:108–115. https://doi.org/10.1016/j.tibtech.2012.12.003
Fernandez-Valades-Gamez R et al. (2016) Usefulness of a bioengineered oral mucosa model for preventing palate bone alterations in rabbits with a mucoperiostial defect. Biomed Mater 11:015015. https://doi.org/10.1088/1748-6041/11/1/015015
Garcia-Martinez L et al (2017) Encapsulation of human elastic cartilage-derived chondrocytes in nanostructured fibrin-agarose hydrogels. Histochem Cell Biol 147:83–95. https://doi.org/10.1007/s00418-016-1485-9
Garzon I et al (2012a) A combined approach for the assessment of cell viability and cell functionality of human fibrochondrocytes for use in tissue engineering. PloS One 7:e51961. https://doi.org/10.1371/journal.pone.0051961
Garzon I et al (2012b) Evaluation of the cell viability of human Wharton’s jelly stem cells for use in cell therapy. Tissue Eng Part C Methods 18:408–419. https://doi.org/10.1089/ten.TEC.2011.0508
Garzon I et al (2014a) Expression of epithelial markers by human umbilical cord stem cells. A topographical analysis. Placenta 35:994–1000. https://doi.org/10.1016/j.placenta.2014.09.007
Garzon I et al (2014b) Generation of a biomimetic human artificial cornea model using Wharton’s jelly mesenchymal stem cells. Investig Ophthalmol Vis Sci 55:4073–4083. https://doi.org/10.1167/iovs.14-14304
Godoy-Guzman C, Nunez C, Orihuela P, Campos A, Carriel V (2018) Distribution of extracellular matrix molecules in human uterine tubes during the menstrual cycle: a histological and immunohistochemical analysis. J Anat. https://doi.org/10.1111/joa.12814
Horman SR et al (2017) Functional profiling of microtumors to identify cancer associated fibroblast-derived drug targets. Oncotarget 8:99913–99930. https://doi.org/10.18632/oncotarget.21915
Jaimes-Parra BD et al (2017) Membranes derived from human umbilical cord Wharton’s jelly stem cells as novel bioengineered tissue-like constructs. Histol Histopathol. https://doi.org/10.14670/HH-11-897
Kalaszczynska I, Ferdyn K (2015) Wharton’s jelly derived mesenchymal stem cells: future of regenerative medicine? Recent findings and clinical significance. BioMed Res Int 2015:430847. https://doi.org/10.1155/2015/430847
Kiernan JA (2008) Histological and histochemical methods: theory and practice, 4th edn. Scion, Oxford
Krotz SP, Robins JC, Ferruccio TM, Moore R, Steinhoff MM, Morgan JR, Carson S (2010) In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary. J Assist Reprod Genet 27:743–750. https://doi.org/10.1007/s10815-010-9468-6
Martin-Piedra MA, Garzon I, Oliveira AC, Alfonso-Rodriguez CA, Carriel V, Scionti G, Alaminos M (2014) Cell viability and proliferation capability of long-term human dental pulp stem cell cultures. Cytotherapy 16:266–277. https://doi.org/10.1016/j.jcyt.2013.10.016
Mills SE (2012) Histology for pathologists/editor: Stacey E. Mills, 4th edn. edn. Lippincott Williams & Wilkins, Philadelphia
Napolitano AP, Chai P, Dean DM, Morgan JR (2007a) Dynamics of the self-assembly of complex cellular aggregates on micromolded nonadhesive hydrogels. Tissue Eng 13:2087–2094. https://doi.org/10.1089/ten.2006.0190
Napolitano AP et al. (2007b) Scaffold-free three-dimensional cell culture utilizing micromolded nonadhesive hydrogels. BioTechniques 43:494, 496–500
Philips C, Cornelissen M, Carriel V (2018) Evaluation methods as quality control in the generation of decellularized peripheral nerve allografts. J Neural Eng 15:021003. https://doi.org/10.1088/1741-2552/aaa21a
Potier E, Rivron NC, Van Blitterswijk CA, Ito K (2016) Micro-aggregates do not influence bone marrow stromal cell chondrogenesis. J Tissue Eng Regen Med 10:1021–1032. https://doi.org/10.1002/term.1887
Rodriguez-Arco L et al (2016) Biocompatible magnetic core-shell nanocomposites for engineered magnetic tissues. Nanoscale 8:8138–8150. https://doi.org/10.1039/c6nr00224b
Roosens A, Puype I, Cornelissen R (2017) Scaffold-free high throughput generation of quiescent valvular microtissues. J Mol Cell Cardiol 106:45–54. https://doi.org/10.1016/j.yjmcc.2017.03.004
Rozario T, DeSimone DW (2010) The extracellular matrix in development and morphogenesis: a dynamic view. Dev Biol 341:126–140. https://doi.org/10.1016/j.ydbio.2009.10.026
Schugar RC et al (2009) High harvest yield, high expansion, and phenotype stability of CD146 mesenchymal stromal cells from whole primitive human umbilical cord tissue. J Biomed Biotechnol 2009:789526. https://doi.org/10.1155/2009/789526
Shen YQ et al. (2018) Combination of melatonin and rapamycin for head and neck cancer therapy: suppression of AKT/mTOR pathway activation, and activation of mitophagy and apoptosis via mitochondrial function regulation J Pineal Res. https://doi.org/10.1111/jpi.12461
Song W et al (2016) Dynamic self-organization of microwell-aggregated cellular mixtures. Soft Matter 12:5739–5746. https://doi.org/10.1039/c6sm00456c
Taghizadeh RR, Cetrulo KJ, Cetrulo CL (2011) Wharton’s Jelly stem cells: future clinical applications. Placenta 32(Suppl 4):S311–S315. https://doi.org/10.1016/j.placenta.2011.06.010
Talaei-Khozani T, Borhani-Haghighi M, Ayatollahi M, Vojdani Z (2015) An in vitro model for hepatocyte-like cell differentiation from Wharton’s jelly derived-mesenchymal stem cells by cell-base aggregates Gastroenterol Hepatol Bed Bench 8:188–199
Williams DF (2014) The biomaterials conundrum in tissue engineering. Tissue Eng Part A 20:1129–1131. https://doi.org/10.1089/ten.tea.2013.0769
Zorlutuna P et al (2012) Microfabricated biomaterials for engineering 3D tissues. Adv Mater 24:1782–1804. https://doi.org/10.1002/adma.201104631
Acknowledgements
This study was supported by the Spanish plan Nacional de investigación Científica, Desarrollo e Innovación Tecnológica, Ministry of Economy and Competitiveness (Instituto de Salud Carlos III), Grant Nos. FIS PI17/393 and FIS PI17/391, co-financed by Fondo Europeo de Desarrollo Regional (ERDF-FEDER, EU), UE and by the Regional Ministry of Health, Junta de Andalucía, Spain, Grant Nos. SAS PI-400-2016 and CS PI-0257-2017. Authors are grateful to Dr. Ariane Ruyffelaert for the editing language service. Daniel Durand was supported by “Consejo Nacional de Ciencia y Tecnología” (CONACYT), scholarship program from Mexican government 580680/694261. Finally, this study forms part of the Doctoral Thesis of Daniel Durand Herrera.
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Durand-Herrera, D., Campos, F., Jaimes-Parra, B.D. et al. Wharton’s jelly-derived mesenchymal cells as a new source for the generation of microtissues for tissue engineering applications. Histochem Cell Biol 150, 379–393 (2018). https://doi.org/10.1007/s00418-018-1685-6
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DOI: https://doi.org/10.1007/s00418-018-1685-6